直接靶向STAT3的苍耳亭通过抑制TSLP释放和NK2细胞极化缓解过敏性气道炎症
2026/03/04
摘要
背景：苍耳亭是一种从传统中药苍耳子中提取的天然倍半萜内酯，据报道具有多种药理活性。尽管苍耳亭抗哮喘的生物活性已得到公认，但其潜在机制尚缺乏深入研究。
目的：系统研究苍耳亭改善过敏性气道炎症的治疗潜力及其机制。
方法：通过为期24天的屋尘螨诱导的小鼠哮喘模型评估苍耳亭的缓解能力。通过检测不同浓度氯化乙酰甲胆碱刺激下的增强呼气间歇值来评估气道反应性。采用酶联免疫吸附法检测血清中的IgE，以及肺组织中IL-4、IL-5、IL-13、IL-25、IL-33、胸腺基质淋巴细胞生成素和IFN-γ的蛋白水平。肺组织切片经苏木精-伊红染色后评估组织学变化。通过流式细胞术分析小鼠外周血及人NK92MI细胞系中自然杀伤细胞亚群的变化。应用抗去唾液酸GM1抗体清除NK细胞，并通过过继转移确定NK2细胞在哮喘中的关键作用。为验证TSLP在诱导NK2极化中的关键作用，使用了抗TSLP单克隆抗体。应用STAT3抑制剂Stattic，并结合流式细胞术和蛋白质印迹法，分析STAT3激活是否是TSLP诱导NK2极化以及支气管上皮细胞产生TSLP不可或缺的环节。采用分子对接、微量热泳动和表面等离子体共振实验检测苍耳亭与STAT3的直接结合。
结果：给予苍耳亭显著缓解了哮喘的典型特征，包括气道高反应性、嗜酸性粒细胞增多、血清IgE升高、气道Th2细胞因子水平升高以及肺组织病理改变。流式细胞术分析显示，苍耳亭特异性减少了肺中产生IL-13的NK2细胞的聚集，但不影响IFN-γ阳性的NK1细胞。清除NK细胞和过继转移NK2细胞证实了NK2细胞在驱动病理过程及介导苍耳亭保护作用中的关键角色。机制上，苍耳亭在体内和体外均选择性抑制了支气管上皮细胞来源的TSLP。
结论：我们确定TSLP是STAT3依赖性NK2细胞分化的新型诱导因子，而苍耳亭可抑制此过程。此外，苍耳亭能以高亲和力直接结合STAT3蛋白。因此，苍耳亭抑制了支气管上皮细胞（抑制TSLP产生）和NK细胞（削弱TSLP诱导的NK2分化）中的STAT3磷酸化。
Direct targeting of STAT3 by xanthatin suppresses allergic airway inflammation via inhibition of TSLP release and NK2 cell polarization
K. F. Bao, J. Zheng, X. T. Wang, Y. J. Zhou, Y. Y. Chen, W. Y. Yuan, et al.
Abstract
BACKGROUND:
Xanthatin is a natural sesquiterpene lactone derived from Xanthium strumarium L. (cang-er-zi) in traditional Chinese medicine (TCM), which has been reported with multiple pharmacological activities. Although the bioactivity of xanthatin against asthma is well recognized, the underlying mechanism lacks thorough research.
OBJECTIVE:
To systematically investigate the therapeutic potential and mechanism of xanthatin ameliorating allergic airway inflammation.
METHODS:
The alleviating capacity of xanthatin was evaluated on a 24-day house dust mite (HDM)- induced mice asthma model. Airway responsiveness was determined by detecting enhanced pause (penh) upon stimulation with different concentrations of methacholine chloride. IgE in serum, and protein levels of IL-4, IL-5, IL-13, IL-25, IL-33, thymic stromal lymphopoietin (TSLP) and IFN-γ in lungs were detected by enzyme-linked immunosorbent assay (ELISA). Lung histological changes were assessed after tissue slices were stained with haematoxylin and eosin (H&E). Changes of subsets of natural killer cells (NK) in mice peripheral blood and human NK92MI cell line were analyzed by flow cytometry. Anti-Asialo GM1 antibody was applied for the depletion of NK, and the key role of NK2 involved in asthma was determined by adoptive transfer. To validate the pivotal role of TSLP in inducing NK2 polarization, anti-TSLP monoclonal antibody was used. The signal trans ducer and activator of transcription 3 (STAT3) inhibitor stattic, alongside with flow cytometry and western blotting, was applied to analyze whether STAT3 activation was the indispensable part for TSLP to induce NK2 and bronchial epithelial cell to produce TSLP. Molecular docking, microscale thermophoresis (MST), and surface plasmon resonance (SPR) assays were applied to test the direct binding of xanthatin to STAT3.
RESULTS:
Xanthatin administration significantly alleviated hallmark asthmatic features, including airway hyper responsiveness, eosinophilia, elevated serum IgE and high levels of Th2 cytokines in airway, and lung tissue pathological changes. Flow cytometry analysis revealed that xanthatin specifically reduced the lung accumula tion of IL-13-producing NK2, but not IFN-γ + NK1. Depletion of NK cells and adoptive transfer of NK2 cells confirmed the pivotal role of NK2 cells in driving the pathology and in mediating xanthatin's protective effects. Mechanistically, xanthatin selectively suppressed bronchial epithelial cell-derived TSLP both in vivo and in vitro.
CONCLUSION:
We identified TSLP as a novel inducer of STAT3-dependent NK2 cell differentiation, which was inhibited by xanthatin. Furthermore, xanthatin directly bound to STAT3 protein with high affinity. Consequently, xanthatin inhibited STAT3 phosphorylation in both bronchial epithelial cells (suppressing TSLP production) and in NK cells (impairing TSLP-induced NK2 differentiation).
背景：苍耳亭是一种从传统中药苍耳子中提取的天然倍半萜内酯，据报道具有多种药理活性。尽管苍耳亭抗哮喘的生物活性已得到公认，但其潜在机制尚缺乏深入研究。
目的：系统研究苍耳亭改善过敏性气道炎症的治疗潜力及其机制。
方法：通过为期24天的屋尘螨诱导的小鼠哮喘模型评估苍耳亭的缓解能力。通过检测不同浓度氯化乙酰甲胆碱刺激下的增强呼气间歇值来评估气道反应性。采用酶联免疫吸附法检测血清中的IgE，以及肺组织中IL-4、IL-5、IL-13、IL-25、IL-33、胸腺基质淋巴细胞生成素和IFN-γ的蛋白水平。肺组织切片经苏木精-伊红染色后评估组织学变化。通过流式细胞术分析小鼠外周血及人NK92MI细胞系中自然杀伤细胞亚群的变化。应用抗去唾液酸GM1抗体清除NK细胞，并通过过继转移确定NK2细胞在哮喘中的关键作用。为验证TSLP在诱导NK2极化中的关键作用，使用了抗TSLP单克隆抗体。应用STAT3抑制剂Stattic，并结合流式细胞术和蛋白质印迹法，分析STAT3激活是否是TSLP诱导NK2极化以及支气管上皮细胞产生TSLP不可或缺的环节。采用分子对接、微量热泳动和表面等离子体共振实验检测苍耳亭与STAT3的直接结合。
结果：给予苍耳亭显著缓解了哮喘的典型特征，包括气道高反应性、嗜酸性粒细胞增多、血清IgE升高、气道Th2细胞因子水平升高以及肺组织病理改变。流式细胞术分析显示，苍耳亭特异性减少了肺中产生IL-13的NK2细胞的聚集，但不影响IFN-γ阳性的NK1细胞。清除NK细胞和过继转移NK2细胞证实了NK2细胞在驱动病理过程及介导苍耳亭保护作用中的关键角色。机制上，苍耳亭在体内和体外均选择性抑制了支气管上皮细胞来源的TSLP。
结论：我们确定TSLP是STAT3依赖性NK2细胞分化的新型诱导因子，而苍耳亭可抑制此过程。此外，苍耳亭能以高亲和力直接结合STAT3蛋白。因此，苍耳亭抑制了支气管上皮细胞（抑制TSLP产生）和NK细胞（削弱TSLP诱导的NK2分化）中的STAT3磷酸化。
（中日友好医院呼吸与危重症医学科 李春晓 摘译 林江涛 审校）
（Journal of Ethnopharmacology 2026 Vol. 362 DOI: 10.1016/j.jep.2026.121296）
（Journal of Ethnopharmacology 2026 Vol. 362 DOI: 10.1016/j.jep.2026.121296）
Direct targeting of STAT3 by xanthatin suppresses allergic airway inflammation via inhibition of TSLP release and NK2 cell polarization
K. F. Bao, J. Zheng, X. T. Wang, Y. J. Zhou, Y. Y. Chen, W. Y. Yuan, et al.
Abstract
BACKGROUND:
Xanthatin is a natural sesquiterpene lactone derived from Xanthium strumarium L. (cang-er-zi) in traditional Chinese medicine (TCM), which has been reported with multiple pharmacological activities. Although the bioactivity of xanthatin against asthma is well recognized, the underlying mechanism lacks thorough research.
OBJECTIVE:
To systematically investigate the therapeutic potential and mechanism of xanthatin ameliorating allergic airway inflammation.
METHODS:
The alleviating capacity of xanthatin was evaluated on a 24-day house dust mite (HDM)- induced mice asthma model. Airway responsiveness was determined by detecting enhanced pause (penh) upon stimulation with different concentrations of methacholine chloride. IgE in serum, and protein levels of IL-4, IL-5, IL-13, IL-25, IL-33, thymic stromal lymphopoietin (TSLP) and IFN-γ in lungs were detected by enzyme-linked immunosorbent assay (ELISA). Lung histological changes were assessed after tissue slices were stained with haematoxylin and eosin (H&E). Changes of subsets of natural killer cells (NK) in mice peripheral blood and human NK92MI cell line were analyzed by flow cytometry. Anti-Asialo GM1 antibody was applied for the depletion of NK, and the key role of NK2 involved in asthma was determined by adoptive transfer. To validate the pivotal role of TSLP in inducing NK2 polarization, anti-TSLP monoclonal antibody was used. The signal trans ducer and activator of transcription 3 (STAT3) inhibitor stattic, alongside with flow cytometry and western blotting, was applied to analyze whether STAT3 activation was the indispensable part for TSLP to induce NK2 and bronchial epithelial cell to produce TSLP. Molecular docking, microscale thermophoresis (MST), and surface plasmon resonance (SPR) assays were applied to test the direct binding of xanthatin to STAT3.
RESULTS:
Xanthatin administration significantly alleviated hallmark asthmatic features, including airway hyper responsiveness, eosinophilia, elevated serum IgE and high levels of Th2 cytokines in airway, and lung tissue pathological changes. Flow cytometry analysis revealed that xanthatin specifically reduced the lung accumula tion of IL-13-producing NK2, but not IFN-γ + NK1. Depletion of NK cells and adoptive transfer of NK2 cells confirmed the pivotal role of NK2 cells in driving the pathology and in mediating xanthatin's protective effects. Mechanistically, xanthatin selectively suppressed bronchial epithelial cell-derived TSLP both in vivo and in vitro.
CONCLUSION:
We identified TSLP as a novel inducer of STAT3-dependent NK2 cell differentiation, which was inhibited by xanthatin. Furthermore, xanthatin directly bound to STAT3 protein with high affinity. Consequently, xanthatin inhibited STAT3 phosphorylation in both bronchial epithelial cells (suppressing TSLP production) and in NK cells (impairing TSLP-induced NK2 differentiation).
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